JP2003137789A - Nitrogen monoxide eliminator - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a nitrogen monoxide eliminator that has high safety to human bodies and can effectively eliminate nitrogen monoxide. SOLUTION: This nitrogen monoxide eliminator contains, as an active ingredient, at least one selected from glucosamine, chitosan oligosaccharides with a polymerization degree of 2-10, low-molecular-weight chitosan with a polymerization degree of 11-100 and their salts. The nitrogen monoxide eliminator is preferably used by adding to at least one selected from drinks, foods, medicines or cosmetics. Particularly, in the cases that glucosamine and/or salts thereof are used as an active ingredient, they may be used as one component for tobacco filers, air cleaner filters, air conditioner filters and car mufflers.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a highly safe nitric oxide scavenger containing glucosamine or the like obtained from shells of crustaceans as an active ingredient.

[0002]

2. Description of the Related Art Nitric oxide (hereinafter referred to as NO) is a nitrogen oxide (NO) which has been a problem as an air pollutant.
x), which is oxidized in the atmosphere to produce nitrogen dioxide (N
O ₂ ). Nitrogen dioxide is said to be a cause of acid rain and chemical oxidants, in addition to affecting respiratory organs at high concentrations. It has also been pointed out that NO in the air may act as an oxidant and attack unsaturated fatty acids contained in sebum to cause rough skin (Yoichi Okubo et al., Journal of Air Pollution Society, 25 (3), 212-220, 19
90).

The cause of generation of NO is mainly due to combustion of fossil fuel such as petroleum and is emitted from factories, automobiles, households and the like. It is also known to be present in cigarette smoke as a more familiar example (Shinagawa, K.
et al: Biochem. Biophys. Res. Commun, 253, 99-10
3, 1998).

On the other hand, NO is known to exist in the living body. NO in vivo (hereinafter referred to as NO in vivo)
Is produced by NO synthase (hereinafter referred to as NOS) using L-arginine, NADPH, tetrahydrobiopterin, calmodulin and the like as substrates, and functions as an extremely important physiologically active molecule. For example, the true identity of vascular endothelium-derived relaxing factor (EDRF) is NO, and it is known that it plays a role of a foreign substance attack in a signal transmitter in the nervous system and in the immune system.

The action of NO in the living body has two sides, and if the amount produced is too large or too small, it will lead to diseases. For example,
Sepsis is a shock state (septic shock) due to a rapid decrease in blood pressure due to the expansion of blood vessels due to excessive NO production.
Is known to cause. In addition, excessively produced NO reacts with active oxygen produced by macrophages to become a strong oxidant, which may cause nerve cell damage and arteriosclerosis, or destroy immune cells themselves such as macrophages. It has been pointed out.

[0006] As a means of reducing the excessively produced in vivo NO, an inhibitor that inactivates NOS (hereinafter referred to as NOS
Inhibitors) and scavengers that react rapidly with NO (hereinafter referred to as NO scavengers) can be used (Tetsuhiko Yoshimura: "From NO (nitric oxide) universe to cells", p.11.
3-134, Kyoritsu Shuppan (1998)). Such NOS inhibitors, N ^omega - monomethyl -L- arginine, N ^omega -
Dimethyl -L- ^arginine, N ω - L- arginine derivative nitro -L- arginine and aminoguanidine or S
-Arginine analogs such as ethylisothiourea. Further, examples of the NO eliminator include an iron complex of diethylenetriaminepentaacetic acid, an iron complex of desferrioxamine, and hydroxocobalamin.

[0007]

However, in clinical trials in which the above-mentioned NOS inhibitor is administered, although the symptoms are improved, there are various causes such as an increase in blood pressure, a decrease in blood flow in organs, and a disorder. It turns out that there is a problem of causing various side effects. Further, the NO scavenger as described above may remove even NO that maintains physiological homeostasis, and its application to living organisms has been unknown.

Therefore, an object of the present invention is to provide a nitric oxide scavenger which is highly safe for the human body and can effectively remove nitric oxide.

[0009]

[Means for Solving the Problem] In the process of studying physiological activities of glucosamine, chitosan oligosaccharides, low molecular weight chitosan, etc., the present inventors found that glucosamine and chitosan oligosaccharides have strong nitric oxide scavenging ability. The present invention has been completed based on this finding.

That is, the nitric oxide scavenger of the present invention is
It is characterized by containing at least one selected from glucosamine, chitosan oligosaccharide having a degree of polymerization of 2 to 10, low molecular weight chitosan having a degree of polymerization of 11 to 100, and salts thereof as an active ingredient.

According to the present invention, by incorporating glucosamine, chitosan oligosaccharide, low molecular weight chitosan, etc., which are naturally derived ingredients, as an active ingredient, nitric oxide can be effectively removed. It is possible to provide a highly safe nitric oxide scavenger without side effects even when added to and ingested.

Since the nitric oxide scavenger of the present invention is water-soluble even in the neutral pH range, it can be easily mixed uniformly with other raw materials when added to foods and drinks, pharmaceuticals, etc. Highly absorbed by the body when ingested.

The nitric oxide scavenger of the present invention is preferably used by adding it to one selected from foods and drinks, pharmaceuticals, or cosmetics. In the case of food and drink, it is particularly effective for food and drink containing a nitrate- or nitrite-based color former or preservative.

Ingestion or administration of foods and drinks or pharmaceuticals to which this nitric oxide scavenger is added can safely and effectively remove excess NO in the living body. For example, septic shock, diabetic complications, and nerve cell disorders. It can be expected to have effects such as prevention of inflammation, bronchial asthma and the like, improvement of these symptoms, and reduction of side effects when taking a remedy for angina and impotence.

Further, nitrates and nitrites added to foods and drinks as color formers and preservatives are reduced to NO by the action of bacteria and react with amines (amino acids) in the foods and drinks to cause carcinogenicity. It is known that nitrosamine, which is a substance, is produced, but by adding the nitric oxide scavenger of the present invention, a nitrosamine production preventing effect can be expected.

Further, by applying the cosmetic containing the nitric oxide scavenger to the skin, the oxidative modification of the skin due to NO in the air can be suppressed, and the effect of preventing rough skin, dullness, etc. can be expected.

Among the nitric oxide scavengers of the present invention, those containing glucosamine and / or a salt thereof as an active ingredient may be selected from filters for cigarettes, filters for air purifiers, filters for air conditioners or filters for mufflers of automobiles. It is preferably used as one selected component.

The above-mentioned filter to which the nitric oxide scavenger is added can effectively reduce the amount of NO contained in cigarette smoke, air or exhaust gas, and therefore adversely affects the health of NO in the air. Can be reduced.

[0019]

BEST MODE FOR CARRYING OUT THE INVENTION Glucosamine, which is an active ingredient of the nitric oxide scavenger of the present invention, is a natural amino sugar distributed in biological components such as glycoproteins, glycolipids and mucopolysaccharides.
Industrially, it can be prepared by hydrolyzing chitin contained in cartilage of crustaceans such as crab, shrimp and krill and cartilage of squid with an acid or an enzyme. Further, glucosamine may be in a free form or may form a salt. Examples of the glucosamine salt include hydrochloride, sulfate and the like. As these glucosamine and salts thereof, commercially available products can be used.

Chitosan oligosaccharides having a degree of polymerization of 2 to 10, which are the active ingredients of the nitric oxide scavenger of the present invention, and a degree of polymerization of 11 to 1
The low molecular weight chitosan of 00 can be prepared by hydrolyzing chitosan obtained by deacetylating chitin with an acid or an enzyme. The chitosan oligosaccharide and low molecular weight chitosan may be in the free form or may form a salt. Examples of salts of chitosan oligosaccharides and low molecular weight chitosan include, for example, hydrochloride, lactate, acetate, sulfate, citrate, ascorbate, malate, succinate, adipate, gluconate, Examples thereof include tartrate salts.

Further, chitosan oligosaccharides and low molecular weight chitosan having the above-mentioned degree of polymerization have high absorbability into the body,
It is expected that NO in vivo can be effectively eliminated when taken orally. Further, since it is water-soluble even in the neutral pH range, it can be easily and uniformly mixed with other raw materials when it is added to foods and drinks, pharmaceuticals, etc., and has excellent processability. Commercially available chitosan oligosaccharides, low molecular weight chitosan, and salts thereof can be used.

Glucosamine, a chitosan oligosaccharide having a degree of polymerization of 2 to 10, a degree of polymerization of 1 in the nitric oxide scavenger of the present invention.
The content of at least one selected from low molecular weight chitosan of 1 to 100 and salts thereof (hereinafter referred to as an active ingredient) is preferably 0.1% by mass or more, more preferably 10% by mass or more, and 20% by mass. The above is particularly preferable.

Glucosamine, a component derived from natural products,
Chitosan oligosaccharides and low molecular weight chitosan have been widely used as raw materials for health foods and pharmaceuticals, and are known to be safe substances with few side effects.

The nitric oxide scavenger of the present invention may appropriately contain excipients, minerals, vitamins, sugars, flavors and the like in addition to the above-mentioned active ingredients. In addition, the product form is
It is not particularly limited, and examples thereof include tablets, powders, granules, solutions, capsules and the like.

The nitric oxide scavenger of the present invention can be used by adding it to one selected from foods and drinks, pharmaceuticals, and cosmetics.

For example, when it is used for food and drink, the amount added is 0.1 to 100 per day in terms of the above-mentioned active ingredients.
The amount that can be ingested by 0 mg is preferable, and the amount that can be ingested by 10 to 1000 mg is more preferable.

When used for pharmaceuticals, the effective dose for oral administration is 0.1 to 1000 mg, more preferably 0.1 to 100 mg, calculated as the above-mentioned active ingredient per day. In the case of intravenous injection and intramuscular injection, 0.01 mg to 1 in terms of the above-mentioned active ingredient per day.
It is 00 mg, more preferably 0.1 to 10 mg.

It is expected that excessive in vivo NO can be safely and effectively eliminated by ingesting or administering the food or drink or the drug to which the nitric oxide eliminating agent of the present invention is added. Therefore, it is necessary to prevent diseases such as septic shock, diabetic complications, nerve cell disorders, inflammation, bronchial asthma and the like caused by excessive production of NO in the body, improve these symptoms, and treat angina and impotence. Effects such as side effects when taking the drug can be expected.

Further, the nitric oxide scavenger of the present invention may be added to foods and drinks (eg, ham, salami, sausages, bacon, tarako, sashiko etc.) containing a nitrate or nitrite type color former or preservative. Therefore, an effect of preventing the generation of nitrosamine can be expected.

When it is used for cosmetics, its addition amount is preferably 0.01 to 10% by mass, more preferably 0.01 to 1.0% by mass in terms of the above-mentioned active ingredient.

By using the cosmetics to which the nitric oxide scavenger of the present invention is added, oxidative denaturation of unsaturated fatty acids and proteins contained in sebum due to NO in the air is suppressed,
The effect of preventing rough skin and dullness can be expected.

Further, the nitric oxide scavenger of the present invention, when it contains glucosamine and / or its salt as an active ingredient, is selected from a cigarette filter, an air purifier filter, an air conditioner filter or an automobile muffler filter. It can be preferably used as one component. Specifically, the nitric oxide scavenger can be used as a microcapsule as a powder, or by filling it in a filter with an appropriate carrier, or by coating and immobilizing it with a semipermeable polymer.

The cigarette filter in the present invention means a filter which is separately attached to a commercially available cigarette and used, or a filter in a commercially available cigarette with a filter. By using this cigarette filter, the amount of NO in cigarette smoke (suction smoke) can be reduced.

When a cigarette filter is filled with a nitric oxide scavenger, the filling amount per filter is 0.1 to 100 mg in terms of glucosamine and / or its salt.
Is preferred, and 10 to 50 mg is more preferred.

[0035]

EXAMPLES The present invention will be described in more detail with reference to examples.

Example 1 As test substances, glucosamine hydrochloride, chitosan oligosaccharide hydrochloride, N-acetylglucosamine, chitin oligosaccharide, cellooligosaccharide and maltooligosaccharide were used to measure the NO scavenging ability.

Further, 1-hydroxy-2 is used as an NO generating reagent.
-Oxo-3- (N-3-methyl-3-aminopropyl) -3-methy
l-1-triazene (hereinafter referred to as NOC-7, manufactured by Dojindo Co., Ltd.) was used as 2- (4-c) as a spin trap agent for NO.
arboxyphenyl) -4,4,5,5-tetramethylimidazoline-1
-Oxyl-3-oxide sodium salt (hereinafter referred to as C-PTIO, manufactured by Labotech Co., Ltd.) was used, and each test substance was adjusted to a concentration of 2.0% (w / v) with 0.1 M phosphorus. It was dissolved in an acid buffer (pH 7.4) before use.

The NO scavenging ability is measured in the literature (Noda et a
l .: Res. Commun. Mol. Pathol. Pharmacol., 96, 115-
124, 1997).

That is, 140 μL of 0.1 M phosphate buffer (pH 7.4) and 20 μL of test substance solution were placed in a test tube.
And 0.1 mM C-PTIO (phosphate buffer solution) were added and mixed, and then 0.1 mM NOC-7 (0.1 N N
20 μL of a solution of aOH was mixed and immediately a flat quartz cell (trade name “ES-LC12”, manufactured by JEOL Ltd.,
The same shall apply hereinafter) and the electron spin resonance apparatus (hereinafter, ES
R device. The product name "JES-FR30", manufactured by JEOL Ltd., the same applies hereinafter).

Exactly 15 minutes after mixing NOC-7, the amount of NO generated was measured using the signal intensity of C-PTI detected when C-PTIO traps NO as an index. MnO was used as the internal standard substance.

With the C-PTI signal intensity when a phosphate buffer solution was used instead of the test substance solution as 100% (control), the relative amount of NO when each test substance was added was calculated and The NO scavenging ability was investigated. The result is shown in FIG.

From FIG. 1, when neutral sugars such as N-acetylglucosamine, chitin oligosaccharides, cellooligosaccharides, and maltooligosaccharides were added, the NO residual ratio was not much different from the control, and the NO elimination ability was low or not observed at all. On the other hand, when glucosamine hydrochloride or chitosan oligosaccharide hydrochloride was added, 98.5% and 95.8% N, respectively were obtained.
It can be seen that O is eliminated and the amino sugar has a very strong NO eliminating ability.

Example 2 Using crystalline cellulose as a carrier, the respective raw materials were mixed in the following blending ratio, and 0.5 g of this powder was filled in a plastic cylinder (diameter 5 mm × length 80 mm) to prepare a cigarette filter. As a control, a filter filled with crystalline cellulose (carrier) was prepared.

Tobacco filter composition (% by mass) Crystalline cellulose 79.9 Glucosamine hydrochloride 20.0 Activated carbon 0.1 Each of the obtained filters was attached to the mouthpiece of a commercially available cigarette,
The cigarette was ignited while being sucked, and was burned to a length of 3 cm from the ignition point, and the sucked smoke that passed through the filter was collected in each test tube containing the spin trap agent. In addition,
The spin trap agent is N-metyl-D-glucamine-dith.
iocarbamate (hereinafter referred to as MGD. Labtech Co., Ltd.)
(MGD) ₂ -Fe ²⁺ complex prepared by mixing an aqueous solution of iron (II) sulfate. In particular,
50 mM MGD solution (0.1 M phosphate buffer (pH
7.4)) 1.0 mL and 0.2 mL of 50 mM iron (II) sulfate aqueous solution were mixed and used immediately.

After 30 minutes, each of the collected liquids was collected in a flat quartz cell and set in an ESR device, and (MGD) ₂ —Fe ²⁺ —N
The spectral intensity of the O complex was measured. Then, the N in the sucked smoke that has passed through the control filter
The amount of O was set to 100% and the amount of NO in the suction smoke that passed through each filter was calculated. The result is shown in FIG.

From FIG. 2, it can be seen that the amount of NO in the suction smoke that has passed through the filter filled with glucosamine hydrochloride is reduced by about 30% as compared with the control. That is, it was revealed that glucosamine hydrochloride has an effect of reducing NO in cigarette smoke.

Example 3 Tablet confectionery, cosmetics, tablets, and a fat emulsion for intravenous injection using the nitric oxide scavenger of the present invention were produced by the following formulations.

[0048]   Tablet confectionery (per tablet 1.5g)     Glucosamine hydrochloride 400mg     Chitosan oligosaccharide hydrochloride 200mg     Lemon juice powder 200mg     Glucose 50mg     Crystalline cellulose 650mg   Cosmetics (foundation)     Soybean phospholipids 50.0% by mass     Glycerin 10.0% by mass     Glucosamine hydrochloride 5.0 mass%     Seaweed extract 3.0 mass%     Hyaluronic acid 0.5% by mass     UV absorber suitable amount     Remaining amount of water for injection as 100   Tablet (200 mg / tablet)     Crystalline cellulose 140mg     Diethylenetriaminepentaacetic acid-iron complex 10 mg     Glucosamine hydrochloride 50mg   Intravenous fat emulsion     Purified soybean phospholipid 10.0 mass%     Chitosan oligosaccharide 1.0% by mass     D-mannitol 5.0 mass%     Water for injection 84.0% by mass

[0049]

As described above, according to the present invention,
Glucosamine, a natural product-derived component, polymerization degree: 2 to 10
Chitosan oligosaccharide, low molecular weight chitosan having a degree of polymerization of 11 to 100, and salts thereof as an active ingredient, nitric oxide can be effectively removed, and side effects even when ingested It is possible to provide a highly safe nitric oxide scavenger. This nitric oxide scavenger can be suitably used for foods, drinks, cosmetics, pharmaceuticals and the like. In particular, a nitric oxide scavenger containing glucosamine and / or a salt thereof can be preferably used as a component of a filter for tobacco, an air purifier, a filter for an air conditioner, or the like.

[Brief description of drawings]

FIG. 1 is a table showing the test results of NO scavenging ability of each test substance.

FIG. 2 is a table showing the measurement results of NO scavenging ability in cigarette smoke of each filter.

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) A61K 7/02 A61K 31/7016 31/7016 31/702 31/702 31/722 31/722 A61P 3/10 A61P 3/10 9/00 9/00 11/06 11/06 25/00 25/00 29/00 29/00 39/02 39/02 43/00 121 43/00 121 C07H 5/06 // C07H 5 / 06 A23L 2/00 FF term (reference) 4B017 LC03 LK13 4B018 MD41 ME14 4C057 BB02 CC04 DD02 4C083 AA112 AC122 AD201 AD202 AD321 AD332 AD572 CC12 4C086 EA02 EA23 MA02 MA35 MA52 MA66 NA14 ZC37 ZC75

Claims (4)

[Claims] 1. A monoxide containing at least one selected from glucosamine, chitosan oligosaccharides having a degree of polymerization of 2 to 10, low molecular weight chitosan having a degree of polymerization of 11 to 100, and salts thereof as an active ingredient. Nitrogen erasing agent. 2. The nitric oxide scavenger according to claim 1, which is used by adding it to one kind selected from foods and drinks, pharmaceuticals, or cosmetics. 3. The nitric oxide scavenger according to claim 2, which is added to a food or drink containing a nitrate or nitrite type color former or a preservative. 4. A composition containing glucosamine and / or a salt thereof as an active ingredient and used as one kind of component selected from a filter for cigarette, a filter for air purifier, a filter for air conditioner and a filter for muffler of an automobile. Item 2. A nitric oxide scavenger according to Item 1.